Figure 3
RFX5 is a positive regulator of KDM4A expression in HCC. (A) KDM4A expression levels were assessed in FLAG-RFX5-transfected HepG2 cells via QRT-PCR analysis. (B) KDM4A expression levels were assessed in shRNA targeting RFX5-transfected HepG2 cells via QRT-PCR analysis. (C) KDM4A protein expression levels were determined in MHCC-97H and HepG2 cells transduced with lentiviral FLAG-RFX5 or sgRNA targeting RFX5 via Western blot analysis. (D) The correlation between RFX5 and KDM4A mRNA expressions in HCC patients derived from TCGA LIHC dataset. (E) The correlation of KDM4A protein expression status determined by IHC analysis with overall survival time (OST). (F) The correlation of KDM4A mRNA expression level determined by RNAseq in TCGA LIHC dataset with OST. (G) Immunohistochemistry analysis was used to quantify the amount of KDM4A protein in HCC tumor tissues and adjacent non-tumor tissues was determined (N−, less than 5% of tumor cells stained positive; N+, 5–30% of tumor cells stained positive; N++, 31–50% of tumor cells stained positive; N+++, > 51% of tumor cells stained positive.). (H) IHC staining of RFX5 and KDM4A in the same cancer nest. (I) The co-expression of RFX5 and KDM4A in HCC tissues was determined by the immunofluorescence staining. The white arrows pointed HCC cells which co-expressed RFX5 and KDM4A.
